Therapeutic combination of a VEGF antagonist and anti-hypertensive agent

ABSTRACT

Disclosed are compositions and methods for treating a disease or condition related to angiogenesis with a vascular endothelial growth factor (VEGF) inhibitor and one or more anti-hypertensive agent(s). The method of the invention is useful for preventing the development of hypertension and/or reducing hypertension in a subject treated with a VEGF inhibitor.

CROSS-REFERENCE TO RELATED APPLICATIONS

This application claims the benefit under 35 USC § 119(e) of U.S.Provisional 60/652,394 filed 11 Feb. 2005, which application is hereinspecifically incorporated by reference in its entirety.

BACKGROUND

1. Field of the Invention

The field of the invention is related to therapeutic methods of treatingdiseases in a mammal with a vascular endothelial growth factor (VEGF)antagonist in combination with one or more anti-hypertensive agents.

2. Description of Related Art

Vascular endothelial growth factor (VEGF) has been recognized as aprimary stimulus of angiogenesis in pathological conditions. Approachesto methods of blocking VEGF include soluble receptor constructs,antisense molecules, RNA aptamers, and antibodies. See, for example, PCTWO/0075319, for a description of VEGF-receptor based antagonists.

Combination therapies using an anti-VEGF antibody and chemotherapeuticagents, such as paclitaxel (TAXOL™), are known (see, for example, U.S.Pat. No. 6,342,219).

BRIEF SUMMARY OF THE INVENTION

In one aspect, the invention features a pharmaceutical compositioncomprising a high affinity vascular endothelial cell growth factor(VEGF) antagonist, one or more anti-hypertensive therapeutic agent(s),and a pharmaceutically acceptable carrier.

More specifically, the VEGF antagonist a high affinity fusion proteindimer (or “trap”) comprising a fusion polypeptide having animmunoglobulin-like (Ig) domain 2 of the VEGF receptor Flt1 and Igdomain 3 of the VEGF receptor Flk1 or Flt4, and a multimerizingcomponent. Even more specifically, the VEGF antagonist comprises afusion polypeptide selected from the group consisting ofFlt1D2.Flk1D3.FcΔC1(a) (SEQ ID NOs:1-2), VEGFR1R2-FcΔC1 (a) (SEQ IDNOs:3-4), or a functionally equivalent thereof.

In specific embodiments, the one or more anti-hypertensive therapeuticagent are selected from the group consisting of ACE inhibitors(ACCUPRIL™ (Parke-Davis); ALTACE™ (Monarch); CAPTOPRIL™ (Mylan);ENALAPRILATE™ (Baxter); LOTENSIN™ (Novartis); MAVIK™ (Bristol-MyersSquibb); PRINIVIL™ (Merck); UNIVASC™ (Schwarz), VASOTEC™ (Merck);calcium-channel antagonists such as nifedipine, β-adrenergic receptorantagonists, such as for example, propanalol, sotalol; angiotensin IIreceptor antagonists; α-adrenergic receptor antagonists; direct activevasodilators; and diuretic agents used in the treatment of hypertension.In preferred embodiments, the anti-hypertensive therapeutic agent is anACE inhibitor or a β-adrenergic receptor blocker.

In a second aspect, the invention features a pharmaceutical compositioncomprising a vascular endothelial cell growth factor (VEGF) antagonist,one or more anti-hypertensive therapeutic agent(s), and apharmaceutically acceptable carrier, wherein the VEGF antagonist is adimer composed of two fusion proteins each having an immunoglobulin-like(Ig) domain 2 of the VEGF receptor Flt1 and Ig domain 3 of the VEGFreceptor Flk1 or Flt4, and a multimerizing component. In specificembodiments, the VEGF antagonist is selected from the group consistingof Flt1D2.Flk1D3.FcΔC1(a) (SEQ ID NOs:1-2), VEGFR1R2-FcΔC1(a) (SEQ IDNOs:3-4), or a functionally equivalent thereof.

In a third aspect, the invention features a method of treating a diseaseor condition which is ameliorated, inhibited, or reduced by a VEGFantagonist in a human, comprising administering a combination of avascular endothelial growth factor (VEGF) antagonist and at least oneanti-hypertensive agent.

The combined therapeutics of the invention achieves maximalanti-angiogenic activity while minimizing the known side effectsresulting from treatment with anti-angiogenic agents, specifically,hypertension. The combination of an anti-angiogenic agent with an ACEinhibitor or angiotensin receptor blocker may also be used to preventproteinuria in subjects at risk thereof.

Diseases and/or conditions, or recurrences thereof, which areameliorated, inhibited, or reduced by treatment with the combination ofthe invention are those treated with a VEGF inhibitor such as the VEGFtrap described above. For example, conditions ameliorated by treatmentwith a VEGF inhibitor include diseases such as cancer or diabetes.Conditions which are ameliorated, inhibited, prevented, or reduced bytreatment with the combined therapeutics of the invention includevascular permeability, edema, or inflammation such as brain edemaassociated with injury, stroke, or tumor, edema associated withinflammatory disorders such as psoriasis or arthritis, asthma, edemaassociated with burns, ascites and pleural effusion associated withtumors, inflammation or trauma, chronic airway inflammation, capillaryleak syndrome, sepsis kidney disease associated with increased leakageof protein, eye disorders such as eye related macular degeneration anddiabetic retinopathy, abnormal angiogenesis such as polycystic ovarydisease, entometriosis and endometrial carcinoma. A VEGF inhibitor mayalso be used to induce regression or reduction of the size of anexisting tumor or metastatic cancer; diabetes, decrease tumorneovascularization, improve transplant corneal survival time, inhibitcorneal transplant rejection or corneal lympangiogenesis andangiogenesis.

A subject to be treated is preferably a subject with one of the abovelisted conditions who suffers from hypertension, is at risk fordevelopment of hypertension or in which the prevention or inhibition ofhypertension is desirable, e.g., a subject at risk for cardiovasculardisease, a subject over 65 years of age, or a patient who cannototherwise be treated with an appropriate dose of the VEGF antagonistwithout developing hypertension.

The VEGF inhibitor and anti-hypertensive agent may be administeredsimultaneously, separately or in combination, or sequentially over arelatively short period of time, e.g., within minutes, hours, or days.

In a fourth aspect, the invention features a method of preventing thedevelopment of hypertension during treatment with a vascular endothelialgrowth factor (VEGF) inhibitor in a patient at risk thereof, comprisingadministering a combination of a VEGF) antagonist and at least oneanti-hypertensive agent.

In a fifth aspect, the invention features a method of treatinghypertension during treatment with a vascular endothelial growth factor(VEGF) inhibitor in a patient at risk thereof, comprising administeringa combination of a VEGF) antagonist and at least one anti-hypertensiveagent.

Other objects and advantages will become apparent from a review of theensuing detailed description.

DETAILED DESCRIPTION

Before the present methods and compositions are described, it is to beunderstood that this invention is not limited to particular methods, andexperimental conditions described, as such methods and conditions mayvary. It is also to be understood that the terminology used herein isfor the purpose of describing particular embodiments only, and is notintended to be limiting, since the scope of the present invention willbe limited only by the appended claims.

As used in this specification and the appended claims, the singularforms “a”, “an”, and “the” include plural references unless the contextclearly dictates otherwise. Thus for example, a reference to “a method”includes one or more methods, and/or steps of the type described hereinand/or which will become apparent to those persons skilled in the artupon reading this disclosure and so forth.

Unless defined otherwise, all technical and scientific terms used hereinhave the same meaning as commonly understood by one of ordinary skill inthe art to which this invention belongs. Although any methods andmaterials similar or equivalent to those described herein can be used inthe practice or testing of the present invention, the preferred methodsand materials are now described. All publications mentioned herein areincorporated herein by reference to describe the methods and/ormaterials in connection with which the publications are cited.

General Description

In the normal mammal, blood pressure is strictly controlled by a complexsystem of physiological factors. This is important for survival becausehigh blood pressure (hypertension) can lead to a number of adversemedical events and conditions, such as, for example, stroke, acutecoronary syndrome, myocardial infarction, and renal failure. Studiesshow that VEGF transiently dilates coronary arteries in vitro (Ku et.al. (1993) Am J Physiol 265:H585-H592) and to induce hypotension (Yanget. al. (1996) J Cardiovasc Pharmacol 27:838-844). Methods for treatingeclampsia and preemclampsia are known (see, for example, US patentapplication publication 2003/0220262, WO 98/28006, WO 00/13703) isdescribed a method for treating hypertension comprising administering toa patient an effective amount of an angiogenic factor such as VEGF, oran agonist thereof. US patent application publication 2003/0144298 showsthat administration of high levels of a VEGF receptor tyrosine kinaseinhibitor leads to a sustained increase in blood pressure in rats whenadministered chronically.

VEGF Antagonists and VEGF-Specific Fusion Polypeptide Traps

In a preferred embodiment, the VEGF antagonist is a dimeric fusionprotein capable of binding VEGF with a high affinity composed of tworeceptor-Fc fusion protein consisting of the principal ligand-bindingportions of the human VEGFR1 and VEGFR2 receptor extracellular domainsfused to the Fc portion of human IgG1 (termed a “VEGF trap”).Specifically, the VEGF trap consists of Ig domain 2 from VEGFR1, whichis fused to Ig domain 3 from VEGFR2, which in turn is fused to the Fcdomain of IgG1 (SEQ ID NO:2).

In a preferred embodiment, an expression plasmid encoding the VEGF trapis transfected into CHO cells, which secrete VEGF trap into the culturemedium. The resulting VEGF trap is a dimeric glycoprotein with a proteinmolecular weight of 97 kDa and contains ˜15% glycosylation to give atotal molecular weight of 115 kDa.

Since the VEGF trap binds its ligands using the binding domains ofhigh-affinity receptors, it has a greater affinity for VEGF than domonoclonal antibodies. The VEGF trap binds VEGF-A (K_(D)=1.5 pM), PLGF1(K_(D)=1.3 nM), and PLGF2 (K_(D)=50 pM); binding to other VEGF familymembers has not yet been fully characterized.

Anti-Hypertensive Therapeutic Agents

The invention may be practiced with a VEGF inhibitor, preferably a VEGFtrap as described in U.S. Pat. No. 6,833,349, herein specificallyincorporated by reference, and an agent which is capable of loweringblood pressure. Anti-hypertensive agents include calcium channelblockers, angiotensin converting enzyme inhibitors (ACE inhibitors),angiotensin II receptor antagonists (A-II antagonists), diuretics,β-adrenergic receptor blockers, vasodilators and α-adrenergic receptorblockers.

Calcium channel blockers include amlodipine; bepridil; clentiazem;diltiazem; fendiline; gallopamil; mibefradil; prenylamine; semotiadil;terodiline; verapamil; aranidipine; barnidipine; benidipine;cilnidipine; efonidipine; elgodipine; felodipine; isradipine;lacidipine; lercanidipine; manidipine; nicardipine; nifedipine;nilvadipine; nimodipine; nisoldipine; nitrendipine; cinnarizine;flunarizine; lidoflazine; lomerizine; bencyclane; etafenone; andperhexiline.

Angiotensin converting enzyme inhibitors (ACE-inhibitors) includealacepril; benazepril; captopril; ceronapril; delapril; enalapril;fosinopril; imidapril; lisinopril; moveltipril; perindopril; quinapril;ramipril; spirapril; temocapril; and trandolapril.

Angiotensin-II receptor antagonists include, but are not limited to:candesartan (U.S. Pat. No. 5,196,444); eprosartan; irbesartan; losartan;and valsartan.

β-blockers include, but are not limited to: acebutolol; alprenolol;amosulalol; arotinolol; atenolol; befunolol; betaxolol; bevantolot;bisoprolol; bopindolol; bucumolol; bufetolol; bufuralol; bunitrolol;bupranolol; butidrine hydrochloride; butofilolol; carazolol; carteolol;carvedilol; celiprolol; cetamolol; cloranololdilevalol; epanolol;indenolol; labetalol; levobunolol; mepindolol; metipranolol; metoprolol;moprolol; nadolol; nadoxolol; nebivalol; nipradilol; oxprenolol;penbutolol; pindolol; practolol; pronethalol; propranolol; sotalol;sulfinalol; talinolol; tertatolol; tilisolol; timolol; toliprolol; andxibenolol.

α-blockers include, but are not limited to: amosulalol; arotinolol;dapiprazole; doxazosin; fenspiride; indoramin; labetolol, naftopidil;nicergoline; prazosin; tamsulosin; tolazoline; trimazosin; andyohimbine.

Vasodilators include cerebral vasodilators, coronary vasodilators andperipheral vasodilators. Cerebral vasodilators include bencyclane;cinnarizine; citicoline; cyclandelate; ciclonicate; diisopropylaminedichloroacetate; eburnamonine; fasudil; fenoxedil; flunarizine;ibudilast; ifenprodil; lomerizine; nafronyl; nicametate; nicergoline;nimodipine; papaverine; tinofedrine; vincamine; vinpocetine; andviquidil.

Coronary vasodilators include, but are not limited to: amotriphene;bendazol; benfurodil hemisuccinate; benziodarone; chloracizine;chromonar; clobenfural; clonitrate; cloricromen; dilazep; dipyridamole;droprenilamine; efloxate; erythrityl tetranitrate; etafenone; fendiline;floredil; ganglefene; hexestrol bis(β-diethylaminoethyl) ether;hexobendine; itramin tosylate; khellin; lidoflazine; mannitolhexanitrate; medibazine; nitroglycerin; pentaerythritol tetranitrate;pentrinitrol; perhexiline; pimefylline; prenylamine; propatyl nitrate;trapidil; tricromyl; trimetazidine; trolnitrate phosphate; visnadine.

Peripheral vasodilators include, but are not limited to: aluminiumnicotinate; bamethan; bencyclane; betahistine; bradykinin; brovincamine;bufeniode; buflomedil; butalamine; cetiedil; ciclonicate; cinepazide;cinnarizine; cyclandelate; diisopropylamine dichloroacetate; eledoisin;fenoxedil; flunarizine; hepronicate; ifenprodil; iloprost; inositolniacinate; isoxsuprine; kallidin; kallikrein; moxisylyte; nafronyl;nicametate; nicergoline; nicofuranose; nylidrin; pentifylline;pentoxifylline; piribedil; prostaglandin E₁; suloctidil; tolazoline; andxanthinol niacinate.

Diuretic includes but is not limited to diuretic benzothiadiazinederivatives, diuretic organomercurials, diuretic purines, diureticsteroids, diuretic sulfonamide derivatives, diuretic uracils and otherdiuretics such as amanozine; amiloride; arbutin; chlorazanil; ethacrynicacid; etozolin; hydracarbazine; isosorbide; mannitol; metochalcone;muzolimine; perhexiline; ticrynafen; triamterene; and urea.

Treatment Population

A human subject preferably treated with the combined therapeuticsdescribed herein is a subject in which it is desirable to prevent orreduce one or more side effects resulting from treatment with ananti-angiogenic agent, such as hypertension, proteinuria. Particularlypreferred subjects are those suffering from hypertension, over 65 yearsof age, or subjects in which reduction of or prevention of undesirableside effects allows a maximal dose of the anti-angiogenic agent to beused which otherwise could not be used without placing the subject atrisk for an adverse medical event. Patients suffering from renal cellcarcinoma, pancreatic carcinoma, advanced breast cancer, colorectalcancer, malignant mesothelioma, multiple myeloma, ovarian cancer, ormelanoma may be treated with the combined therapeutics of the invention.Diseases and/or conditions, or recurrences thereof, which areameliorated, inhibited, or reduced by treatment with the combinedtherapeutics of the invention cancer, diabetes, vascular permeability,edema, or inflammation such as brain edema associated with injury,stroke, or tumor, edema associated with inflammatory disorders such aspsoriasis or arthritis, asthma, edema associated with burns, ascites andpleural effusion associated with tumors, inflammation or trauma, chronicairway inflammation, capillary leak syndrome, sepsis kidney diseaseassociated with increased leakage of protein, eye disorders such as eyerelated macular degeneration and diabetic retinopathy, abnormalangiogenesis such as polycystic ovary disease, entometriosis andendometrial carcinoma. A VEGF inhibitor may also be used to induceregression or reduction of the size of an existing tumor or metastaticcancer; diabetes, decrease tumor neovascularization, improve transplantcorneal survival time, inhibit corneal transplant rejection or corneallympangiogenesis and angiogenesis.

Combination Therapies

In numerous embodiments, a VEGF antagonist may be administered incombination with one or more additional compounds or therapies,including a second VEGF antagonist molecule. Combination therapyincludes administration of a single pharmaceutical dosage formulationwhich contains a VEGF antagonist and one or more additional agents; aswell as administration of a VEGF antagonist and one or more additionalagent(s) in its own separate pharmaceutical dosage formulation. Forexample, a VEGF antagonist and a cytotoxic agent, a chemotherapeuticagent or a growth inhibitory agent can be administered to the patienttogether in a single dosage composition such as a combined formulation,or each agent can be administered in a separate dosage formulation.Where separate dosage formulations are used, the VEGF-specific fusionprotein of the invention and one or more additional agents can beadministered concurrently, or at separately staggered times, i.e.,sequentially.

The term “cytotoxic agent” as used herein refers to a substance thatinhibits or prevents the function of cells and/or causes destruction ofcells. The term is intended to include radioactive isotopes (e.g. I¹³¹,I¹²⁵, Y⁹⁰ and Re¹⁸⁶), chemotherapeutic agents, and toxins such asenzymatically active toxins of bacterial, fungal, plant or animalorigin, or fragments thereof.

A “chemotherapeutic agent” is a chemical compound useful in thetreatment of cancer. Examples of chemotherapeutic agents includealkylating agents such as thiotepa and cyclosphosphamide (CYTOXAN®);alkyl sulfonates such as busulfan, improsulfan and piposulfan;aziridines such as benzodopa, carboquone, meturedopa, and uredopa;ethylenimines and methylamelamines including altretamine,triethylenemelamine, trietylenephosphoramide,triethylenethiophosphaoramide and trimethylolomelamine; nitrogenmustards such as chlorambucil, chlornaphazine, cholophosphamide,estramustine, ifosfamide, mechlorethamine, mechlorethamine oxidehydrochloride, melphalan, novembichin, phenesterine, prednimustine,trofosfamide, uracil mustard; nitrosureas such as carmustine,chlorozotocin, fotemustine, lomustine, nimustine, ranimustine;antibiotics such as aclacinomysins, actinomycin, authramycin, azaserine,bleomycins, cactinomycin, calicheamicin, carabicin, carminomycin,carzinophilin, chromomycins, dactinomycin, daunorubicin, detorubicin,6-diazo-5-oxo-L-norleucine, doxorubicin, epirubicin, esorubicin,idarubicin, marcellomycin, mitomycins, mycophenolic acid, nogalamycin,olivomycins, peplomycin, potfiromycin, puromycin, quelamycin,rodorubicin, streptonigrin, streptozocin, tubercidin, ubenimex,zinostatin, zorubicin; anti-metabolites such as methotrexate and5-fluorouracil; folic acid analogues such as denopterin, methotrexate,pteropterin, trimetrexate; purine analogs such as fludarabine,6-mercaptopurine, thiamiprine, thioguanine; pyrimidine analogs such asancitabine, azacitidine, 6-azauridine, carmofur, cytarabine,dideoxyuridine, doxifluridine, enocitabine, floxuridine; androgens suchas calusterone, dromostanolone propionate, epitiostanol, mepitiostane,testolactone; anti-adrenals such as aminoglutethimide, mitotane,trilostane; folic acid replenisher such as frolinic acid; aceglatone;aldophosphamide glycoside; aminolevulinic acid; amsacrine; bestrabucil;bisantrene; edatraxate; defofamine; demecolcine; diaziquone;elfornithine; elliptinium acetate; etoglucid; gallium nitrate;hydroxyurea; lentinan; lonidamine; mitoguazone; mitoxantrone; mopidamol;nitracrine; pentostatin; phenamet; pirarubicin; podophyllinic acid;2-ethylhydrazide; procarbazine; PSK®; razoxane; sizofiran;spirogermanium; tenuazonic acid; triaziquone;2,2′,2″-trichlorotriethylamine; urethan; vindesine; dacarbazine;mannomustine; mitobronitol; mitolactol; pipobroman; gacytosine;arabinoside (“Ara-C”); cyclophosphamide; thiotepa; taxanes, e.g.paclitaxel (TAXOL®, Bristol-Myers Squibb Oncology, Princeton, N.J.) anddocetaxel (TAXOTERE®; Aventis Antony, France); chlorambucil;gemcitabine; 6-thioguanine; mercaptopurine; methotrexate; platinumanalogs such as cisplatin and carboplatin; vinblastine; platinum;etoposide (VP-16); ifosfamide; mitomycin C; mitoxantrone; vincristine;vinorelbine; navelbine; novantrone; teniposide; daunomycin; aminopterin;xeloda; ibandronate; CPT-11; topoisomerase inhibitor RFS 2000;difluoromethylornithine; retinoic acid; esperamicins; capecitabine; andpharmaceutically acceptable salts, acids or derivatives of any of theabove. Also included in this definition are anti-hormonal agents thatact to regulate or inhibit hormone action on tumors such asanti-estrogens including for example tamoxifen, raloxifene, aromataseinhibiting 4(5)-imidazoles, 4-hydroxytamoxifen, trioxifene, keoxifene,LY 117018, onapristone, and toremifene (Fareston); and anti-androgenssuch as flutamide, nilutamide, bicalutamide, leuprolide, and goserelin;and pharmaceutically acceptable salts, acids or derivatives of any ofthe above.

A “growth inhibitory agent” when used herein refers to a compound orcomposition which inhibits growth of a cell, especially a cancer celleither in vitro or in vivo. Examples of growth inhibitory agents includeagents that block cell cycle progression (at a place other than Sphase), such as agents that induce G1 arrest and M-phase arrest.Classical M-phase blockers include the vincas (vincristine andvinblastine), TAXOL®, and topo II inhibitors such as doxorubicin,epirubicin, daunorubicin, etoposide, and bleomycin. Those agents thatarrest G1 also spill over into S-phase arrest, for example, DNAalkylating agents such as tamoxifen, prednisone, dacarbazine,mechlorethamine, cisplatin, methotrexate, 5-fluorouracil, and ara-C.

Methods of Administration

The invention provides compositions and methods of treatment comprisinga VEGF antagonist, such as a VEGF antagonist, and an anti-hypertensiveagent. Various delivery systems are known and can be used to administerthe composition of the invention, e.g., encapsulation in liposomes,microparticles, microcapsules, recombinant cells capable of expressingthe compound, receptor-mediated endocytosis (see, e.g., Wu and Wu, 1987,J. Biol. Chem. 262:4429-4432), construction of a nucleic acid as part ofa retroviral or other vector, etc. Methods of introduction can beenteral or parenteral and include but are not limited to intradermal,intramuscular, intraperitoneal, intravenous, subcutaneous, intranasal,intraocular, and oral routes. The compounds may be administered by anyconvenient route, for example by infusion or bolus injection, byabsorption through epithelial or mucocutaneous linings (e.g., oralmucosa, rectal and intestinal mucosa, etc.) and may be administeredtogether with other biologically active agents. Administration can besystemic or local. Administration can be acute or chronic (e.g. daily,weekly, monthly, etc.) or in combination with other agents. Pulmonaryadministration can also be employed, e.g., by use of an inhaler ornebulizer, and formulation with an aerosolizing agent.

In another embodiment, the active agent can be delivered in a vesicle,in particular a liposome, in a controlled release system, or in a pump.In another embodiment where the active agent of the invention is anucleic acid encoding a protein, the nucleic acid can be administered invivo to promote expression of its encoded protein, by constructing it aspart of an appropriate nucleic acid expression vector and administeringit so that it becomes intracellular, e.g., by use of a retroviral vector(see, for example, U.S. Pat. No. 4,980,286), by direct injection, or byuse of microparticle bombardment, or coating with lipids or cell-surfacereceptors or transfecting agents, or by administering it in linkage to ahomeobox-like peptide which is known to enter the nucleus.

In a specific embodiment, it may be desirable to administer thepharmaceutical compositions of the invention locally to the area in needof treatment; this may be achieved, for example, and not by way oflimitation, by local infusion during surgery, topical application, e.g.,by injection, by means of a catheter, or by means of an implant, theimplant being of a porous, non-porous, or gelatinous material, includingmembranes, such as silastic membranes, fibers, or commercial skinsubstitutes.

A composition useful in practicing the methods of the invention may be aliquid comprising an agent of the invention in solution, in suspension,or both. The term “solution/suspension” refers to a liquid compositionwhere a first portion of the active agent is present in solution and asecond portion of the active agent is present in particulate form, insuspension in a liquid matrix. A liquid composition also includes a gel.The liquid composition may be aqueous or in the form of an ointment.

An aqueous suspension or solution/suspension useful for practicing themethods of the invention may contain one or more polymers as suspendingagents. Useful polymers include water-soluble polymers such ascellulosic polymers and water-insoluble polymers such as cross-linkedcarboxyl-containing polymers. An aqueous suspension orsolution/suspension of the present invention is preferably viscous ormuco-adhesive, or even more preferably, both viscous and mucoadhesive.

Pharmaceutical Compositions

The present invention provides pharmaceutical compositions comprising aVEGF antagonist, an anti-hypertensive agent, and a pharmaceuticallyacceptable carrier. The term “pharmaceutically acceptable” meansapproved by a regulatory agency of the Federal or a state government orlisted in the U.S. Pharmacopeia or other generally recognizedpharmacopeia for use in animals, and more particularly, in humans. Theterm “carrier” refers to a diluent, adjuvant, excipient, or vehicle withwhich the therapeutic is administered. Such pharmaceutical carriers canbe sterile liquids, such as water and oils, including those ofpetroleum, animal, vegetable or synthetic origin, such as peanut oil,soybean oil, mineral oil, sesame oil and the like. Suitablepharmaceutical excipients include starch, glucose, lactose, sucrose,gelatin, malt, rice, flour, chalk, silica gel, sodium stearate, glycerolmonostearate, talc, sodium chloride, dried skim milk, glycerol,propylene, glycol, water, ethanol and the like. The composition, ifdesired, can also contain minor amounts of wetting or emulsifyingagents, or pH buffering agents. These compositions can take the form ofsolutions, suspensions, emulsion, tablets, pills, capsules, powders,sustained-release formulations and the like. Examples of suitablepharmaceutical carriers are described in “Remington's PharmaceuticalSciences” by E. W. Martin.

The composition of the invention can be formulated as neutral or saltforms. Pharmaceutically acceptable salts include those formed with freeamino groups such as those derived from hydrochloric, phosphoric,acetic, oxalic, tartaric acids, etc., and those formed with freecarboxyl groups such as those derived from sodium, potassium, ammonium,calcium, ferric hydroxides, isopropylamine, triethylamine, 2-ethylaminoethanol, histidine, procaine, etc.

The amount of the composition of the invention that will be effectivefor its intended therapeutic use can be determined by standard clinicaltechniques based on the present description. In addition, in vitroassays may optionally be employed to help identify optimal dosageranges. Generally, suitable dosage ranges for intravenous administrationare generally about 20-500 micrograms of active compound per kilogrambody weight. Suitable dosage ranges for intranasal administration aregenerally about 0.01 pg/kg body weight to 1 mg/kg body weight. Effectivedoses may be extrapolated from dose-response curves derived from invitro or animal model test systems.

For systemic administration, a therapeutically effective dose can beestimated initially from in vitro assays. For example, a dose can beformulated in animal models to achieve a circulating concentration rangethat includes the IC₅₀ as determined in cell culture. Such informationcan be used to more accurately determine useful doses in humans. Initialdosages can also be estimated from in vivo data, e.g., animal models,using techniques that are well known in the art. One having ordinaryskill in the art could readily optimize administration to humans basedon animal data.

Dosage amount and interval may be adjusted individually to provideplasma levels of the compounds that are sufficient to maintaintherapeutic effect. In cases of local administration or selectiveuptake, the effective local concentration of the compounds may not berelated to plasma concentration. One having skill in the art will beable to optimize therapeutically effective local dosages without undueexperimentation.

The amount of compound administered will, of course, be dependent on thesubject being treated, on the subject's weight, the severity of theaffliction, the manner of administration, and the judgment of theprescribing physician. The therapy may be repeated intermittently whilesymptoms are detectable or even when they are not detectable. Thetherapy may be provided alone or in combination with other drugs.

Kits

The invention also provides an article of manufacturing comprisingpackaging material and a pharmaceutical agent contained within thepackaging material, wherein the pharmaceutical agent comprises at leastone VEGF antagonist and at least one anti-hypertensive agent, andwherein the packaging material comprises a label or package insert whichindicates that the VEGF antagonist and anti-hypertensive agent can beused for treating cancer or reducing tumor growth.

Other features of the invention will become apparent in the course ofthe following descriptions of exemplary embodiments which are given forillustration of the invention and are not intended to be limitingthereof.

EXAMPLES

The following example is put forth so as to provide those of ordinaryskill in the art with a complete disclosure and description of how tomake and use the methods and compositions of the invention, and are notintended to limit the scope of what the inventors regard as theirinvention. Efforts have been made to ensure accuracy with respect tonumbers used (e.g., amounts, temperature, etc.) but some experimentalerrors and deviations should be accounted for. Unless indicatedotherwise, parts are parts by weight, molecular weight is averagemolecular weight, temperature is in degrees Centigrade, and pressure isat or near atmospheric.

Example 1 Treatment of Malignant Pleural Effusion and Prevention ofHypertension

Adult patients with pathologic diagnosis of stage IIIB-IV NSCLC who areeligible for systemic chemotherapy, and also have an MPE which requirestherapeutic drainage are eligible for inclusion in the study. Patientsundergoing chemotherapy with another agent, or having prior chemotherapywith an inhibitor of VEGF, or active or untreated brain metastases areexcluded. Treatment with VEGF antagonist (SEQ ID NO:4) is an intravenousdose of 300-5000 mg/kg and an anti-hypertensive agent such as an ACEinhibitor or a β-adrenergic receptor blocker. The anti-hypertensivetherapeutic agent may be given separately or in combination with theVEGF antagonist, prior to administration of the VEGF antagonist,simultaneously, or following administration of the VEGF antagonist.

Example 2 Treatment of Solid Tumor and Prevention of Hypertension

Patients with refractory solid tumors or non-Hodgkin's lymphomareceiving no concurrent treatment for their cancer are treated with theVEGF trap (SEQ ID NO:4) as follows. The dose levels range from 100 to5000 mg/kg given subcutaneously. Each patient receives a single initialdose of the VEGF trap followed by weekly injections at the required doselevel. Blood pressure is monitored and tumor burden is assessed at thebeginning and end of the weekly dosing period; patients with stabledisease, partial or complete responses may continue dosing for up to anadditional 6 months in a continuation study.

1. A pharmaceutical composition comprising a high affinity vascularendothelial cell growth factor (VEGF) antagonist, one or moreanti-hypertensive therapeutic agent(s), and a pharmaceuticallyacceptable carrier, wherein the VEGF antagonist is VEGFR1R2-FcΔC1(a)(SEQ ID NO:4).
 2. The pharmaceutical composition of claim 1, wherein theone or more anti-hypertensive therapeutic agent is(are) selected fromthe group consisting of an angiotensin converting enzyme (ACE)inhibitor, a calcium-channel antagonist, a β-adrenergic receptorantagonist, an angiotensin II receptor antagonist, an α-adrenergicreceptor antagonist, a vasodilator and a diuretic agent.
 3. Thepharmaceutical composition of claim 2, wherein the ACE inhibitor isselected from the group consisting of alacepril; benazepril; captopril;ceronapril; delapril; enalapril; fosinopril; imidapril; lisinopril;moveltipril; perindopril; quinapril; ramipril; spirapril; temocapril;and trandolapril.
 4. The pharmaceutical composition of claim 2, whereinthe calcium-channel blocker is selected from the group consisting ofamlodipine, bepridil, clentiazem, diltiazem, fendiline, gallopamil,mibefradil, prenylamine, semotiadil, terodiline, verapamil, aranidipine,barnidipine, benidipine, cilnidipine, efonidipine, elgodipine,felodipine, isradipine, lacidipine, lercanidipine, manidipine,nicardipine, nifedipine, nilvadipine, nimodipine, nisoldipine,nitrendipine, cinnarizine, flunarizine, lidoflazine, lomerizine,bencyclane, etafenone, and perhexiline.
 5. The pharmaceuticalcomposition of claim 2, wherein the β-adrenergic receptor antagonist isselected from the group consisting of acebutolol, alprenolol,amosulalol, arotinolol, atenolol, befunolol, betaxolol, bevantolot,bisoprolol, bopindolol, bucumolol, bufetolol, bufuralol, bunitrolol,bupranolol, butidrine hydrochloride, butofilolol, carazolol, carteolol,carvedilol, celiprolol, cetamolol, cloranololdilevalol, epanolol,indenolol, labetalol, levobunolol, mepindolol, metipranolol, metoprolol,moprolol, nadolol, nadoxolol, nebivalol, nipradilol, oxprenolol,penbutolol, pindolol, practolol, pronethalol, propranolol, sotalol,sulfinalol, talinolol, tertatolol, tilisolol, timolol, toliprolol, andxibenolol.
 6. The pharmaceutical composition of claim 2, wherein theα-adrenergic receptor antagonist is selected from the group consistingof amosulalol, arotinolol, dapiprazole, doxazosin, fenspiride,indoramin, labetolol, naftopidil, nicergoline, prazosin, tamsulosin,tolazoline, trimazosin, and yohimbine.
 7. The pharmaceutical compositionof claim 2, wherein the vasodilator is a cerebral vasodilator, acoronary vasodilator, and/or a peripheral vasodilator.
 8. Thepharmaceutical composition of claim 7, wherein the cerebral vasodilatoris selected from the group consisting of bencyclane, cinnarizine,citicoline, cyclandelate, ciclonicate, diisopropylamine dichloroacetate,eburnamonine, fasudil, fenoxedil, flunarizine, ibudilast, ifenprodil,lomerizine, nafronyl, nicametate, nicergoline, nimodipine, papaverine,tinofedrine, vincamine, vinpocetine, and viquidil.
 9. The pharmaceuticalcomposition of claim 7, wherein the coronary vasodilator is selectedfrom the group consisting of amotriphene, bendazol, benfurodilhemisuccinate, benziodarone, chloracizine, chromonar, clobenfural,clonitrate, cloricromen, dilazep, dipyridamole, droprenilamine,efloxate, erythrityl tetranitrate, etafenone, fendiline, floredil,ganglefene, hexestrol bis(β-diethylaminoethyl) ether, hexobendine,itramin tosylate, khellin, lidoflazine, mannitol hexanitrate,medibazine, nitroglycerin, pentaerythritol tetranitrate, pentrinitrol,perhexiline, pimefylline, prenylamine, propatyl nitrate, trapidil,tricromyl, trimetazidine, trolnitrate phosphate, visnadine.
 10. Thepharmaceutical composition of claim 7, wherein the peripheralvasodilator is selected from the group consisting of aluminiumnicotinate, bamethan, bencyclane, betahistine, bradykinin, brovincamine,bufeniode, buflomedil, butalamine, cetiedil, ciclonicate, cinepazide,cinnarizine, cyclandelate, diisopropylamine dichloroacetate, eledoisin,fenoxedil, flunarizine, hepronicate, ifenprodil, iloprost, inositolniacinate, isoxsuprine, kallidin, kallikrein, moxisylyte, nafronyl,nicametate, nicergoline, nicofuranose, nylidrin, pentifylline,pentoxifylline, piribedil, prostaglandin E₁, suloctidil, tolazoline, andxanthinol niacinate.
 11. The pharmaceutical composition of claim 2,wherein diuretic agent is selected from the group consisting ofbenzothiadiazine derivatives, diuretic organomercurials, diureticpurines, diuretic steroids, diuretic sulfonamide derivatives, diureticuracils, amanozine, amiloride, arbutin, chlorazanil, ethacrynic acid,etozolin, hydracarbazine, isosorbide, mannitol, metochalcone,muzolimine, perhexiline, ticrynafen, triamterene, and urea.